Specificity determinants revealed by the structure of glycosyltransferase Campylobacter concisusPglA
Nemanja Vuksanovic,
Jozlyn R. Clasman,
Barbara Imperiali
et al.
Abstract:In selected Campylobacter species, the biosynthesis of N‐linked glycoconjugates via the pgl pathway is essential for pathogenicity and survival. However, most of the membrane‐associated GT‐B fold glycosyltransferases responsible for diversifying glycans in this pathway have not been structurally characterized which hinders the understanding of the structural factors that govern substrate specificity and prediction of resulting glycan composition. Herein, we report the 1.8 Å resolution structure of Campylobacte… Show more
“…The flexibility along with the hydrophilicity/hydrophobicity of the loop seem to be the most important factors for the donor specificity of UGTs in this region. Lastly, it is worth noting that a loop very similar to the UGT N5 loop is observed in prokaryotic GT-B fold GTs, where structural and mutational studies indicate similar importance in the donor specificity arguing for evolutionary conservation and potentially functional importance ( Liu B. et al, 2021 ; Vuksanovic et al, 2024 ).…”
Section: A Loop In the N-terminal Domain Can Significantly Alter Acti...mentioning
Plant family 1 glycosyltransferases (UGTs) represent a formidable tool to produce valuable natural and novel glycosides. Their regio- and stereo-specific one-step glycosylation mechanism along with their inherent wide acceptor scope are desirable traits in biotechnology. However, their donor scope and specificity are not well understood. Since different sugars have different properties in vivo and in vitro, the ability to easily glycodiversify target acceptors is desired, and this depends on our improved understanding of the donor binding site. In the aim to unlock the full potential of UGTs, studies have attempted to elucidate the structure-function relationship governing their donor specificity. These efforts have revealed a complex phenomenon, and general principles valid for multiple enzymes are elusive. Here, we review the studies of UGT donor specificity, and attempt to group the information into key concepts which can help shape future research. We zoom in on the family-defining PSPG motif, on two loop residues reported to interact with the C6 position of the sugar, and on the role of active site arginines in donor specificity. We continue to discuss attempts to alter and expand the donor specificity by enzyme engineering, and finally discuss future research directions.
“…The flexibility along with the hydrophilicity/hydrophobicity of the loop seem to be the most important factors for the donor specificity of UGTs in this region. Lastly, it is worth noting that a loop very similar to the UGT N5 loop is observed in prokaryotic GT-B fold GTs, where structural and mutational studies indicate similar importance in the donor specificity arguing for evolutionary conservation and potentially functional importance ( Liu B. et al, 2021 ; Vuksanovic et al, 2024 ).…”
Section: A Loop In the N-terminal Domain Can Significantly Alter Acti...mentioning
Plant family 1 glycosyltransferases (UGTs) represent a formidable tool to produce valuable natural and novel glycosides. Their regio- and stereo-specific one-step glycosylation mechanism along with their inherent wide acceptor scope are desirable traits in biotechnology. However, their donor scope and specificity are not well understood. Since different sugars have different properties in vivo and in vitro, the ability to easily glycodiversify target acceptors is desired, and this depends on our improved understanding of the donor binding site. In the aim to unlock the full potential of UGTs, studies have attempted to elucidate the structure-function relationship governing their donor specificity. These efforts have revealed a complex phenomenon, and general principles valid for multiple enzymes are elusive. Here, we review the studies of UGT donor specificity, and attempt to group the information into key concepts which can help shape future research. We zoom in on the family-defining PSPG motif, on two loop residues reported to interact with the C6 position of the sugar, and on the role of active site arginines in donor specificity. We continue to discuss attempts to alter and expand the donor specificity by enzyme engineering, and finally discuss future research directions.
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